A SIMPLE PERFUSION TECHNIQUE FOR ISOLATION OF MATERNAL INTERVILLOUS BLOOD MONONUCLEAR-CELLS FROM HUMAN PLACENTAE

A noninvasive perfusion method for the recovery of maternal placental (intervillous) blood for use in immunologic assays is described. 60% o f the perfused blood samples tested for fetal red blood cell (RBC) con tamination were found to be pure maternal blood; in the remainder, fet al RBC contamination, with a single exception, was less than 6%. The i ntervillous mononuclear cells (IVBMC) isolated from this blood were of predominantly maternal origin as demonstrated by a polymerase chain r eaction-based DNA typing technique. The number of IVBMC obtained was w ithin the range of 9 to 55 x 10(6) cells. Phenotypic analysis of IVBMC surface antigens revealed that 61% of the cells were CD3 + T-cells an d 18% were CD19 + B-cells. The CD4 + and CD8 + T-lymphocyte subsets ac counted for 28 and 26% of the IVBMC, respectively. The IVBMC were func tionally competent as evidenced by in vitro lymphoproliferation and cy tokine production in response to mitogen and PPD stimulation. This tec hnique allows for rapid and safe isolation of large numbers of IVBMC w hich are functionally active up to 12 h post-delivery, thus representi ng a significant improvement over previously described methods. It sho uld facilitate more vigorous research in the study of uteroplacental i mmunity and infectious disease research, particularly in field setting s where sample collection and laboratory facilities are distant. (C) 1 997 Elsevier Science B.V.